Fig 2: Impacts of XST and Cef on the levels of GLT1, AQP4, and MMP-9 in APP/PS1 mice. (A) Sample images of WB analysis of brain AQP4 and GLT1 levels after treatment with XST in APP/PS1 mice. (B) Densitometric analysis of protein levels of AQP4 (p = 0.1113, t-test, n = 4 mice per group) and GLT1 (p = 0.0035, t-test, n = 4 mice per group) after treatment with XST, normalized against GAPDH. (C) Sample images of AQP4 staining in cortex (S1) of APP/PS1 mice after treatment with XST for 15 days. Scale bar, 50 µm. (D) Quantification of AQP4 polarity after XST treatment (F (1, 16) = 183.4, C57 vs. AD, p < 0.0001; AD-Veh vs. AD-XST, p = 0.0073, two-way ANOVA with Bonferroni’s multiple comparisons, n = 5 mice per group). (E) Sample images of WB analysis of brain AQP4 and GLT1 levels after treatment with Cef in APP/PS1 mice. (F) Densitometric analysis of AQP4 (p = 0.1113, t-test, n = 4 mice per group) and GLT1 (p = 0.035, t-test, n = 4 mice per group) levels after treatment with Cef, normalized to GAPDH. (G) Sample images of AQP4 in the cortex (S1) after treatment with Cef for 15 days in APP/PS1 mice. Scale bar, 50 µm (low). Scale bar, 10 µm (high). (H) Quantification of AQP4 polarity after treatment with Cef for 15 days (p = 0.855, t-test, n = 4 mice per group). (I) Sample Western blots of brain MMP-9 levels after treatment with either XST (upper) or Cef (lower) in APP/PS1 mice. (J) Normalized levels of MMP-9 after treatment with XST (p = 0.003, t-test, n = 3 mice per group) or Cef (p = 0.390, t-test, n = 3 mice per group). (K) Quantitative measurement of VEGF-C levels in serum in WT and APP/PS1 mice (p = 0.6928, t-test, n = 6 mice per group). Data are mean ± SEM. *p < 0.05; **p < 0.01.

Fig 3: MiR-483-3p alleviates impairments of learning ability, memory ability, and neural apoptosis in homocysteine-treated rats. (a) The efficacy of AAV-miR-483-3p in hippocampus of rats was detected by PCR. (b) The escape latency of rats in sham group, AAV-miR-483-3p, Hcy+NC group, and Hcy+AAV-miR-483-3p group during the training was recorded. (c), (d) The swimming track of rats during the training and in MWM test. (e) The number of platform crossings after removing the platform. (f), (g) The escape latency and time of rats spent in the target quadrant in MWM test. (h) The percentage of freezing after footshock. (i) Fear conditioning test for contextual fear memory assessment in rats. (j) The protein levels of APP, BACE1 and Aß1-42 in hippocampus of rats were analyzed by western blot. (k) TUNEL assay was conducted to assess apoptosis in hippocampus of rats in the sham, AAV-miR-483-3p, Hcy+NC, and Hcy+AAV-miR-483-3p groups. (l) Protein levels of apoptosis markers (Bcl-2, Bax, and cleaved caspase-3) in hippocampus of rats were quantified using western blot. n = 10/group. * p < .05, ** p < .01 compared with sham group; & p < .05, && p < .01 compared with Hcy group

Fig 4: Effect of electroacupuncture at the Baihui (DU20) acupoint on the expression and processing of BDNF. (A) Representative western blot demonstrating protein expression levels of BDNF and proBDNF in the hippocampus from each group. (B) Ratio of BDNF to proBDNF normalized to the WT group. (C) Representative western blot indicating the levels of TrkB and p-TrkB. (D) Ratio of p-TrkB vs. total TrkB normalized to the WT group. (E) Representative western blot demonstrating the levels of p75NTR in each group. (F) Expression levels of p75NTR normalized to the WT group. **P<0.01 vs. the WT group; #P<0.05, ##P<0.01 and ?P>0.05 vs. the APP/PS1 group. Each group, n=5. BDNF, brain-derived neurotrophic factor; WT, wild-type; NA, non-acupoint; TrkB, tropomyosin receptor kinase B; p75NTR, p75 neurotrophin receptor; p, phosphorylated.

Fig 5: SOD activity (A) and MDA concentration (B) in the hippocampus of male WT and APP/PS1 mice fed a control or high iron diet at age 30 wk. Values are means ± SDs, n = 7. APP/PS1, amyloid precursor protein/presenilin 1; APP/PS1-Ctrl, APP/PS1 mice fed a control diet; APP/PS1-High Fe, APP/PS1 mice fed a high iron diet; MDA, malondialdehyde; SOD, superoxide dismutase; WT, wild-type; WT-Ctrl, wild-type mice fed a control diet; WT-High Fe, wild-type mice fed a high iron diet.